The present invention relates to methods of treatment for irritable bowel syndrome and for nonulcer dyspepsia.
2,3-Benzodiazepines
Certain 2,3-benzodiazepines have been explored extensively for their potent CNS modulating activity. Compounds such as tofisopam (Grandaxin(copyright))(structure shown below, with the atom numbering system indicated), girisopam, and norisopam have demonstrated substantial anxiolytic and antipsychotic activity. 
Tofisopam has been shown in humans to have an activity profile that is significantly different from that of widely used 1,4-benzodiazepine (BZ) anxiolytics such as diazepam (Valium(copyright)) and chlordiazepepoxide (Librium(copyright)). The 1,4-benzodiazepines, in addition to having sedative-hypnotic activity, also possess muscle relaxant and anticonvulsant properties which, though therapeutically useful in some disease states, are nonetheless potentially untoward side effects. Thus the 1,4-benzodiazepines, though safe when administered alone, may be dangerous in combination with other CNS drugs, including alcohol.
Tofisopam, in contrast, is a non-sedative anxiolytic that has no appreciable sedative, muscle relaxant or anticonvulsant properties See Horvath el al., Progress in Neurobiology, 60 (2000), 309-342, the entire disclosure of which is incorporated herein by reference. In clinical studies, tofisopam improved rather than impaired psychomotor performance and showed no interaction with ethanol (Id.). These observations comport with data that show that tofisopam does not interact with central BZ receptors and binds only weakly to peripheral BZ receptors.
Other 2,3-benzodiazepines that are structurally similar to tofisopam have been investigated and shown to have varying activity profiles. For example, GYKI-52466 and GYKI-53655 (structures shown below) act as noncompetitive glutamate antagonists at the AMPA (xcex1-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid) site, and have demonstrated neuroprotective, muscle relaxant and anticonvulsant activity (Id.). Another group of 2,3-benzodiazepines that have been investigated are represented by the compound GYKI-52895, and show activity as selective dopamine uptake inhibitors with potential use in antidepressant and anti-Parkinsonism therapy. 
Tofisopam is a racemic mixture of (R)- and (S)-enantiomers. This is due to the asymmetric carbon, i.e., a carbon with four different groups attached, at the 5-position of the benzodiazepine ring.
The structure and conformational properties of tofisopam have been determined by NMR, CD and x-ray crystallography See Visy el al., Chirality 1:271-275 (1989), the entire disclosure of which is incorporated herein by reference. The 2,3-diazepine ring exists as two different conformers. The major conformers, (+)R and (xe2x88x92)S have the 5-ethyl group in a quasi-equatorial position. In the minor conformers, (xe2x88x92)R and (+)S, the 5-ethyl group is positioned quasi-axially. Thus, racemic tofisopam may exist as four molecular species, i.e., two enantiomers, each of which exists in two conformations. The sign of the optical rotation is reversed upon inversion of the diazepine ring from one conformer to the other. In crystal form, tofisopam exists only as the major conformations, with dextrorotatory tofisopam being of the (R) absolute configuration. See Toth et al., J Heterocyclic Chem., 20:709-713 (1983); Fogassy et al., Bioorganic Heterocycles, Van der Plas, H. C., xc3x96tvxc3x6s, L, Simongi, M., eds. Budapest Amsterdam: Akademia; Kiado-Elsevier, 229:233 (1984), the entire disclosures of which are incorporated herein by reference.
Differential binding of these two conformations of tofisopam is reported in binding studies with human albumin See Simongi et al. Biochem. Pharm., 32(12), 1917-1920, 1983, the entire disclosure of which is incorporated herein by reference. The two conformers have also been reported as existing in equilibrium See Zsila et al., Journal of Liquid Chromatography and Related Technologies, 22(5), 713-719, 1999; and references therein, the entire disclosures of which are incorporated herein by reference.
The optically pure (R)-enantiomer of tofisopam (R)-1-(3,4-dimethoxyphenyl)-4-methyl-5-ethyl-7,8-dimethoxy-5H-2,3-benzodiazepine) has been isolated and shown to possess the nonsedative anxiolytic activity of the racemic mixture. See U.S. Pat. No. 6,080,736; the entire disclosure of which is incorporated herein by reference.
Metabolism of Tofisopam
Tofisopam is metabolized in human, rat, dog, monkey and rabbit to one or more of six major metabolites, depending on the host species:
See Tomori et al., Journal of Chromatography, 241 (1982), p. 89-99.
Of the compounds named above, Compounds 1, 3 and 5 have been identified as metabolites in humans. These compounds have been synthesized and tested in certain pharmacological assays. See C. Ito, xe2x80x9cBehavioral Pharmacological Study on the Structure Activity Relationship of Benzodiazepine Derivatives: With Particular Reference to the Activity of 2,3-Benzodiazepine,xe2x80x9d J. Tokyo Med. College, 39:369-384 (1981).
In an assay of inhibition of aggression in mice, Compound 1 and 3 showed 0% inhibition of aggression and Compound 5 showed a 28.6% inhibition of aggression. In an assay of muricide (mouse killing behavior) in rats, Compound 3 exhibited 0% inhibition of muricide while Compounds 1 and 5 each exhibited a 20% inhibition of muricide. In assays testing for anti-noradrenergic effects, Compound 1 exhibited no effect, while Compounds 3 and 5 demonstrated measurable activity. See Ito, Id.
Compounds 1, 3, 5 and 6 are also disclosed in U.S. Pat. No. 4,322,346, the entire disclosure of which is incorporated herein by reference. Compound 3 is reported therein to demonstrate narcosis-potentiating activity in mice.
Irritable Bowel Syndrome
Irritable bowel syndrome (IBS) is a common disorder that has a pronounced effect on the quality of life and that accounts for a large proportion of healthcare costs. IBS is defined on the basis of the recently modified Rome criteria as (A) the presence for at least 12 weeks (not necessarily consecutive) in the preceding 12 months of abdominal discomfort or pain that cannot be explained by structural or biochemical abnormalities, and (B) at least two of the following three (1) pain relieved with defecation; (2) pain, when the onset thereof is associated with a change in the frequency of bowel movements (diarrhea or constipation); and pain, when the onset thereof is associated with a change in the form of the stool (lose, watery, or pellet-like). IBS may be divided into four subcategories according to whether the predominant symptom is abdominal pain, diarrhea, constipation, or constipation alternating with diarrhea.
Approximately 15 percent of U.S. adults report symptoms that are consistent with the diagnosis of the IBS; the disease affects three times as many women as men. Whether this difference reflects a true predominance of the disorder among women or merely the fact women are more likely to seek medical care has not been determined. IBS is the most common diagnosis made by gastroenterologists in the United States and accounts for 12 percent of visits to primary care providers. It is estimated that only 25 percent of persons with this condition seek medical care for it, and studies suggest that those who seek care are more likely to have behavioral and psychiatric problems than are those who do not seek care. In addition, patients with a diagnosis of IBS are at increased risk for other, non-gastrointestinal functional disorders such as fibromyalgia and interstitial cystitis. The irritable bowel syndrome accounts for an estimate $8 billion in direct medical costs and $25 billion in indirect costs annually in the United States.
Converging evidence supports the concept that IBS results from altered regulation of gastrointestinal motility and epithelial function, as well as altered perception of visceral events. See Mayer et al., Digestive Diseases, 2001, 19:212-218, the entire disclosure of which is incorporated herein by reference.
Altered bowel motility, visceral hypersensitivity, psychosocial factors, an imbalance in neurotransmitters, and infection have all been proposed as playing a part in the development of the irritable bowel syndrome. See B. Horwitz et al., The New England Journal of Medecine, 344:24, 2001, the entire disclosure of which is incorporated herein by reference.
Nonulcer Dyspepsia
Nonulcer dyspepsia (NUD) has been defined as chronic or recurrent upper abdominal pain or discomfort for a period of more than three months"" duration, with symptoms present for more than 25 percent of the time, in the absence of another organic cause. See Fisher R S, Parkman H P, xe2x80x9cManagement of nonulcer dyspepsia,xe2x80x9d New Engl J Med 1998; 339: 1376-1381, the entire disclosure of which is incorporated herein by reference. A simpler definition is that offered by Locke: xe2x80x9cpersistent or recurrent upper abdominal pain or discomfort with no structural or biochemical explanation for the patient""s symptoms.xe2x80x9d See Locke GR, xe2x80x9cNonulcer dyspepsia: what it is and what it is not,xe2x80x9d Mayo Clin Proc 1999;74:1011-15, the entire disclosure of which is incorporated herein by reference
NUD may include bloating, nausea, early satiety, eructation and heartburn. It is a symptom complex rather than a specific condition. An organic cause is found in only 40 percent of patients with dyspepsia. The most common causes are gastroduodenal ulcer, GERD, gastroparesis, and gastric cancer. Other causes include cholelithiasis or choledocolithiasis, pancreatitis, carbohydrate malabsorption, intestinal parasites, NSAID or other medication injury, diabetes, thyroid disorders or connective tissue disorders, ischemic bowel, and abdominal cancer. The 60 percent of patients without an organic cause are considered to have xe2x80x9cnonulcer dyspepsia,xe2x80x9d and fall into a continuum of functional gastrointestinal disorders, including irritable bowel syndrome, functional heartburn, and noncardiac chest pain. See Freidman L S., xe2x80x9cHelicobacter pylori and Nonulcer Dyspepsia,xe2x80x9d New Engl J Med 1998; 339: 1928-30, the entire disclosure of which is incorporated herein by reference.
NUD has many similarities to IBS. NUD and IBS are usually differentiated by whether the abdominal pain is associated with abnormal bowel habits. If the association is present, the condition is considered to be IBS rather than NUD.
Like IBS, the cause of NUD is not well understood. NUD is most likely caused by an alteration in the perception of sensations arising from the gut. Other possible causes of NUD have been investigated, including Helicobacter pylori infection (a bacterial infection in the stomach that is associated with ulcer disease), and an alteration in the function of the stomach, resulting in a delayed emptying of the stomach contents. About 25-50% of patients with NUD have slowed emptying from the stomach which may, in part, explain the increased symptoms after meals.
New agents are needed which are useful in the treatment of IBS and NUD. In particular, agents are needed that are appropriate for chronic long-term use in treatment and prevention these chronic disorders.
In one embodiment of the invention a method of treating or preventing IBS or NUD is provided, comprising administering to an individual in need of such treatment an effective amount of at least one compound according to formula I: 
wherein:
R1 is xe2x80x94(C1-C7)hydrocarbyl or xe2x80x94(C2-C6)heteroalkyl;
R2 is xe2x80x94H or xe2x80x94(C1-C7)hydrocarbyl; wherein R1 and R2 may combine to form a carbocyclic or heterocyclic 5- or 6-membered ring; and
one of R3, R4, R5 or R6 (hereinafter, collectively xe2x80x9cphenyl ring substituentsxe2x80x9d) is xe2x80x94OH, and the remaining phenyl ring substituents are independently selected from the group consisting of xe2x80x94(C1-C7)hydrocarbyl, xe2x80x94CF3, xe2x80x94O(C1-C7)hydrocarbyl, xe2x80x94O-acyl, xe2x80x94NH2, xe2x80x94NH(C1-C6)alkyl, xe2x80x94N((C1-C6)alkyl)2, xe2x80x94NH-acyl and halogen, wherein R5 and R6 may combine to form a 5, 6- or 7-membered heterocyclic ring; and
wherein the carbon-carbon single bond designated by  indicates that the absolute conformation about C* may be either (R) or (S);
or a pharmaceutically acceptable salt thereof.
According to a first sub-embodiment of a compound of formula I, one of R3 or R4 is xe2x80x94OH, and the other phenyl ring substituents are independently selected from the group consisting of xe2x80x94(C1-C7)hydrocarbyl, xe2x80x94CF3, xe2x80x94O(C1-C7)hydrocarbyl, xe2x80x94O-acyl, xe2x80x94NH2, xe2x80x94NH(C1-C6)alkyl, xe2x80x94N((C1-C6)alkyl)2, xe2x80x94NH-acyl and halogen.
According to a second sub-embodiment of a compound of formula I, one of R3 or R4 is xe2x80x94OH, one or two of the other phenyl ring substituents is xe2x80x94OCH3, and the other phenyl ring substituent(s) are independently selected from the group consisting of xe2x80x94(C1-C7)hydrocarbyl, xe2x80x94CF3, xe2x80x94O(C1-C7)hydrocarbyl, xe2x80x94O-acyl, xe2x80x94NH2, xe2x80x94NH(C1-C6)alkyl, xe2x80x94N((C1-C6)alkyl)2, xe2x80x94NH-acyl and halogen.
According to a third sub-embodiment of a compound of formula I, one phenyl ring substituent is xe2x80x94OH, and the other phenyl ring substituents are independently selected from xe2x80x94O(C1-C7)hydrocarbyl.
According to a fourth sub-embodiment of a compound of formula I, one phenyl ring substituent is xe2x80x94OH, and the other phenyl ring substituents are independently selected from xe2x80x94O(C1-C7)alkyl.
According to a fifth sub-embodiment of a compound of formula I, one phenyl ring substituent is xe2x80x94OH, and the other phenyl ring substituents are xe2x80x94OCH3.
Preferably, the phenyl ring substituent which is xe2x80x94OH in the third, fourth and fifth sub-embodiments, is R3 or R4.
In some embodiments of the invention, R1 and R2 are independently selected from xe2x80x94(C1-C7)alkyl, preferably, xe2x80x94(C1-C3)alkyl. In a preferred sub-embodiments R1 is xe2x80x94CH2CH3 and R2 is xe2x80x94CH3.
Preferred compounds for use in the practice of the invention, are selected from the group consisting of:
1-(3,4-dimethoxyphenyl)-4-methyl-5-ethyl-7-hydroxy-8-methoxy-5H-2,3-benzodiazepine;
1-(3-hydroxy-4-methoxyphenyl)-4-methyl-5-ethyl-7,8-dimethoxy-5H-2,3-benzodiazepine;
1-(3-methoxy-4-hydroxyphenyl)-4-methyl-5-ethyl-7,8-dimethoxy-5H-2,3-benzodiazepine;
1-(3,4-dimethoxyphenyl)-4-methyl-5-ethyl-7-methoxy-8-hydroxy-5H-2,3-benzodiazepine;
or pharmaceutically-acceptable salts thereof.
More preferred compounds for use in the practice of the invention, are selected from the group consisting of:
1-(3-hydroxy-4-methoxyphenyl)-4-methyl-5-ethyl-7,8-dimethoxy-5H-2,3-benzodiazepine;
1-(3-methoxy-4-hydroxyphenyl)-4-methyl-5-ethyl-7,8-dimethoxy-5H-2,3-benzodiazepine;
or pharmaceutically-acceptable salts thereof.
According to some embodiments, there are provided the methods above wherein the compound is in the form of the substantially isolated (R)-enantiomer.
According to other embodiments, there are provided the methods above wherein the compound is in the form of the substantially isolated (S)-enantiomer.
The term xe2x80x9cirritable bowel syndromexe2x80x9d refers to a disorder, often recurrent, characterized by abnormally increased motility of the small and large intestines, producing abdominal pain, constipation, or diarrhea.
The term xe2x80x9cnonulcer dyspepsiaxe2x80x9d refers to a disorder manifesting chronic or recurrent upper abdominal pain or discomfort for a period of more than three months"" duration, with symptoms present for more than 25 percent of the time, in the absence of another organic cause.
The term xe2x80x9cacylxe2x80x9d means a radical of the general formula xe2x80x94C(xe2x95x90O)xe2x80x94R, wherein xe2x80x94R is hydrogen, hydrocarbyl, amino, alkylamino, dialkylamino hydroxy or alkoxy.xe2x80x9d Examples include for example, acetyl (xe2x80x94C(xe2x95x90O)CH3), propionyl (xe2x80x94C(xe2x95x90O)CH2CH3), benzoyl (xe2x80x94C(xe2x95x90O)C6H5), phenylacetyl (xe2x80x94C(xe2x95x90O)CH2C6H5), carboethoxy (xe2x80x94CO2CH2CH3), and dimethylcarbamoyl (xe2x80x94C(xe2x95x90O)N(CH3)2). When the R group in the acetyl radical is alkoxy, alkyl amino or dialkyl amino, the alkyl portion is preferably (C1-C6)alkyl, more preferably (C1-C3)alkyl. When the R is hydrocarbyl, it is preferably (C1-C7)hydrocarbyl. When R is hydrocarbyl, it is preferably alkyl, more preferably (C1-C6)alkyl.
The term xe2x80x9calkylxe2x80x9d, by itself or as part of another substituent means, unless otherwise stated, a straight, branched or cyclic chain hydrocarbon radical, including di- and multi-radicals, having the number of carbon atoms designated (i.e. C1-C6 means one to six carbons). Alkyl groups include straight chain, branched chain or cyclic groups, with straight being preferred. Examples include: methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl, cyclohexyl and cyclopropylmethyl. (C1-C6)alkyl is preferred. Most preferred is (C1-C3)alkyl, particularly ethyl, methyl and isopropyl.
The term xe2x80x9calkoxyxe2x80x9d employed alone or in combination with other terms means, unless otherwise stated, an alkyl group having the designated number of carbon atoms, as defined above, connected to the rest of the molecule via an oxygen atom, such as, for example, methoxy, ethoxy, 1-propoxy, 2-propoxy (isopropoxy) and the higher homologs and isomers. Preferred is (C1-C6)alkoxy. More preferred is (C1-C3)alkoxy, particularly ethoxy and methoxy.
The term xe2x80x9caminexe2x80x9d or xe2x80x9caminoxe2x80x9d refers to radicals of the general formula xe2x80x94NRRxe2x80x2, wherein R and Rxe2x80x2 are independently selected from hydrogen or a hydrocarbyl radical, or wherein R and Rxe2x80x2 combined form a heterocycle. Examples of amino groups include: xe2x80x94NH2, methyl amino, diethyl amino, anilino, benzyl amino, piperidinyl, piperazinyl and indolinyl. Preferred hydrocarbyl radicals are (C1-C7)hydrocarbyl radicals. Preferred are hydrocarbyl radicals that are alkyl radicals. More preferred is (C1-C6)alkyl.
The term xe2x80x9caromaticxe2x80x9d refers to a carbocycle or heterocycle having one or more polyunsaturated rings having aromatic character (4n+2) delocalized xcfx80 (pi) electrons).
The term xe2x80x9carylxe2x80x9d employed alone or in combination with other terms, means, unless otherwise stated, a carbocyclic aromatic system containing one or more rings (typically one, two or three rings) wherein such rings may be attached together in a pendent manner, such as a biphenyl, or may be fused, such as naphthalene. Examples include phenyl; anthracyl; and naphthyl.
The term xe2x80x9chydrocarbylxe2x80x9d refers to any moiety comprising only hydrogen and carbon atoms. This definition includes for example alkyl, alkenyl, alkynyl, aryl and benzyl groups. Preferred are (C1-C7)hydrocarbyl.
The term xe2x80x9cheteroalkylxe2x80x9d by itself or in combination with another term means, unless otherwise stated, a stable straight or branched chain radical consisting of the stated number of carbon atoms and one or two heteroatoms selected from the group consisting of O, N, and S. Nitrogen and sulfur atoms may be optionally oxidized to the N-oxide and sulfoxide or sulfone, respectively. In addition, a nitrogen heteroatom may be optionally quaternized. The heteroatom(s) may be placed at any position of the heteroalkyl group, including between the rest of the heteroalkyl group and the fragment to which it is attached, as well as attached to the most distal carbon atom in the heteroalkyl group. Preferred are (C2-C6)heteroalkyl. More preferred are (C2-C4)heteroalkyl. Examples include: xe2x80x94Oxe2x80x94CH2xe2x80x94CH2xe2x80x94CH3, xe2x80x94CH2xe2x80x94CH2CH2xe2x80x94OH, xe2x80x94CH2xe2x80x94CH2xe2x80x94NHxe2x80x94CH3, xe2x80x94CH2xe2x80x94C(xe2x95x90O)xe2x80x94CH3, xe2x80x94CH2xe2x80x94Nxe2x95x90Nxe2x80x94CH2xe2x80x94CH3, xe2x80x94CH2xe2x80x94Sxe2x80x94CH2xe2x80x94CH3, xe2x80x94CH2CH2xe2x80x94S(xe2x95x90O)xe2x80x94CH3 and xe2x80x94CH2xe2x80x94CH2xe2x80x94NHxe2x80x94SO2xe2x80x94CH3. Up to two heteroatoms may be consecutive, such as, for example, xe2x80x94CH2xe2x80x94NHxe2x80x94OCH3, or xe2x80x94CH2xe2x80x94CH2xe2x80x94Sxe2x80x94Sxe2x80x94CH3. More preferred are heteroalkyl groups containing one or two oxygen atoms.
When two groups may xe2x80x9ccombine to form a carbocyclic or heterocyclic 5- or 6-membered ring,xe2x80x9d a carbocyclic ring is preferably saturated. Preferred heterocyclic rings are saturated rings containing one or two heteroatoms selected from N, O and S. Heterocyclic rings annulated to the benzodiazepine seven-membered ring in this way include, for example, furan, dihydrofuran, tetrahydrofuran, pyran, dihydropyran, tetrahydropyran, thiophene, dihydrothiophene, tetrahydrothiophene, pyrrole, dihydropyrrole, pyrrolidine, pyridine, dihydropyridine, tetrahydropyridine and piperidine.
When two groups may xe2x80x9ccombine to form a 5-, 6- or 7-membered heterocyclic ring,xe2x80x9d preferred heterocyclic rings are 5- or 6-membered rings containing one or two heteroatoms selected from N, O and S. More preferred are heterocyclic rings containing one heteroatom selected from N, O and S. Heterocyclic rings annulated to the benzodiazepine phenyl ring in this way include, for example, furan, dihydrofuran, dioxane, dioxolane, pyran, dihydropyran, tetrahydropyran, thiophene, dihydrothiophene, pyridine, dihydropyridine, tetrahydropyridine, piperidine, pyrrole, dihydropyrrole, imidazole, dihydroimidazole, thiazole, dihydrothiazole, oxazole, and dihydrooxazole.
The term xe2x80x9csubstitutedxe2x80x9d means that an atom or group of atoms has replaced hydrogen as the substituent attached to another group. For aryl and heteroaryl groups, the term xe2x80x9csubstitutedxe2x80x9d refers to any level of substitution, namely mono-, di-, tri-, tetra-, or penta-substitution, where such substitution is permitted. The substituents are independently selected, and substitution may be at any chemically accessible position.
The phrase xe2x80x9coptically activexe2x80x9d refers to a property whereby a material rotates the plane of plane-polarized light. A compound that is optically active is nonsuperimposable on its mirror image. The property of nonsuperimposablity of an object on its mirror image is called chirality.
The property of xe2x80x9cchiralityxe2x80x9d in a molecule may arise from any structural feature that makes the molecule nonsuperimposable on its mirror image. The most common structural feature producing chirality is an asymmetric carbon atom, i.e., a carbon atom having four nonequivalent groups attached thereto.
The term xe2x80x9cenantiomerxe2x80x9d refers to each of the two nonsuperimposable isomers of a pure compound that is optically active. Single enantiomers are designated according to the Cahn-Ingold-Prelog system, a set of priority rules that rank the four groups attached to an asymmetric carbon. See March, Advanced Organic Chemistry, 4th Ed., (1992), p. 109. Once the priority ranking of the four groups is determined, the molecule is oriented so that the lowest ranking group is pointed away from the viewer. Then, if the descending rank order of the other groups proceeds clockwise, the molecule is designated (R) and if the descending rank of the other groups proceeds counterclockwise, the molecule is designated (S). In the example below, the Cahn-Ingold-Prelog ranking sequence id A greater than B greater than C greater than D. The lowest ranking atom, D is oriented away from the viewer. 
The term xe2x80x9cracematexe2x80x9d or the phrase xe2x80x9cracemic mixturexe2x80x9d refers to a 50-50 mixture of two enantiomers such that the mixture does not rotate plane-polarized light.
By xe2x80x9c(R)-enantiomer substantially free of the (S)-enantiomerxe2x80x9d is meant a compound of formula I that comprises 80% or more by weight of the (R)-enantiomer and likewise contains 20% or less of the (S)-enantiomer as a contaminant, by weight. Likewise, By xe2x80x9c(S)-enantiomer substantially free of the (R)-enantiomerxe2x80x9d is meant a compound of formula I that comprises 80% or more by weight of the (S)-enantiomer and likewise contains 20% or less of the (R)-enantiomer as a contaminant, by weight.
The term xe2x80x9ceffective amountxe2x80x9d when used to describe the amount of drug administered to a patient suffering from irritable bowel syndrome, refers to the amount of a compound that prevents or alleviates the symptoms of IBS, when administered to a patient suffering from a disorder which manifests chronic or acute symptoms of IBS.
An xe2x80x9ceffective amountxe2x80x9d of the compound when used to describe the amount of drug administered to a patient suffering from nonulcer dyspepsia, refers to the amount of a compound that prevents or alleviates the symptoms of NUD, when administered to a patient suffering from a disorder which manifests chronic or acute symptoms of NUD.
The term xe2x80x9cindividualxe2x80x9d or xe2x80x9csubjectxe2x80x9d includes human beings and non-human animals. With respect to the disclosed methods of treating IBS and NUD, these terms refer, unless the context indicates otherwise, to an organism that is afflicted with or diagnosed with such a disorder.
With respect to disclosed methods of xe2x80x9cpreventingxe2x80x9d or xe2x80x9cdelaying the onsetxe2x80x9d of IBS or NUD, these terms refer unless the context indicates otherwise, to an organism that has a medical history of IBS or NUD that manifests as a recurrent disorder.
According to the present invention, compounds of formula I, and pharmaceutically acceptable salts thereof, are useful in methods of treatment or prevention of irritable bowel syndrome or nonulcer dyspepsia.
Compounds used in the methods of the present invention that comprise substantially isolated enantiomers preferably have a composition that is 85% by weight or greater of the desired enantiomer, and 15% by weight, or less, of the other enantiomer. More preferably, compounds used in methods of the present invention have a composition that is 90% by weight or greater of the desired enantiomer and 10% by weight, or less, of the other enantiomer. More preferably, compounds used in the methods of the present invention have a composition that is 95% by weight or greater of the desired enantiomer and 5% by weight, or less, of the other enantiomer. Most preferably, compounds used in the methods of the present invention have a composition that is 99% by weight or greater of the desired enantiomer and 1% by weight, or less, of the other enantiomer.
The compounds of formula I useful in the methods of the present invention may be prepared by one of several methods. These methods generally follow the synthetic strategies and procedures used in the synthesis of 2,3-benzodiazepines such as tofisopam and tofisopam analogs. See U.S. Pat. Nos. 3,736,315 and 4,423,044 (tofisopam syntheses) and Horvath et al., Progress in Neurobiology 60(2000) p.309-342 and references cited therein (preparation of tofisopam and analogs thereof), the disclosures of which are incorporated herein by reference. In the synthesis methods that follow, the products of the chemical synthesis are racemic compounds of formula I. The racemic mixture may be subsequently separated using known methods of resolution to produce the (R)-enantiomer substantially free of the corresponding (S)-enantiomer, and the (S)-enantiomer substantially free of the corresponding (R)-enantiomer.
2,3-Benzodiazepines of formula I may be synthesized from the corresponding 2-benzopyrilium salt H by reaction with hydrazine hydrate, wherein Xxe2x88x92 is a counterion such as, for example perchlorate: 
Accordingly, hydrazine hydrate (98%, approximately 3 equivalents based on the 2-benzopyrylium salt) is added dropwise to a stirred solution of the 2-benzopyrylium salt H in glacial acetic acid (approximately 1 mL/3 mmol of 2-benzopyrylium salt). During this operation, the solution is maintained at an elevated temperature, preferably, 80-100xc2x0 C. The solution is then maintained a higher elevated temperature, preferably 95-100xc2x0 C., for about one hour. Then the reaction mixture is diluted with 2% aqueous sodium hydroxide solution (approximately 3 equivalents based on the 2-benzopyrylium salt) and cooled. The product 2,3-benzodiazepine separates as a solid and is removed by filtration, washed with water and dried. The crude product may be purified by taking it up in a polar aprotic solvent such as dimethylformamide (DMF) at an elevated temperature, preferably 100-130xc2x0 C., and decolorizing the solution with activated carbon. The carbon is removed by filtration and the filtered solution is diluted with water. The purified product precipitates out of the solution and is collected by filtration.
See Kxc3x3rxc3x3si et al., U.S. Pat. No. 4,322,346, the entire disclosure of which is incorporated herein by reference, disclosing three variations of the reaction protocol for preparing a substituted 2,3-benzodiazepine from the precursor benzopyrilium salt.
Retrosynthetically, the intermediate benzopyrilium salt, H, may be prepared from one of several starting materials. According to one such method, illustrated in Scheme 1, intermediate H is prepared from the corresponding aryl ethanol derivative D via the isochroman intermediate F; wherein Xxe2x88x92 is a counterion such as, for example, perchlorate: 
According to Scheme 1, 3,4-disubstituted ethylbenzoate, A is dissolved in a suitable solvent, preferably ether and cooled to 0xc2x0 C. Two equivalents of a selected Grignard reagent are added dropwise and the reaction is allowed to warm to room temperature and monitored for disappearance of starting material. When the reaction is complete, it may be quenched with a proton source such as acetic acid. Volatiles are removed in vacuo, and the product B is used for the next step without purification.
The xcex1-substituted benzyl alcohol, B, is taken up in a high boiling solvent such as toluene and a catalytic amount of para-toluene sulfonic acid (p-TsOH). The mixture is warmed to reflux and may be monitored for disappearance of starting materials. When the reaction is complete, the volatiles are removed in vacuo and the crude product C is purified by column chromatography.
The substituted styrene, C is hydroxylated under anti-Markovnikov conditions to give intermediate phenylethyl alcohol D. A solution of D, and of a substituted benzaldehyde, E (1.2 eq) are dissolved in anhydrous dioxane. The resulting solution is then saturated with gaseous HCl and warmed, preferably to reflux temperature for about one hour. The mixture is then cooled to room temperature, poured into water, basified, preferably with aqueous sodium hydroxide and extracted with an organic solvent, preferably ethyl acetate. The extract is dried, filtered and concentrated under vacuum. The resulting residue is purified, preferably by crystallization to yield F.
To a stirred, cooled, (preferably to 0-5xc2x0 C.) solution of F (2 g) in acetone (30 mL), is added dropwise a solution of chromium trioxide (2 g) in 35% sulfuric acid (20 mL); added at a rate such that the reaction temperature remains below 5xc2x0 C. After the addition is complete, the reaction mixture is allowed to rise to room temperature and is stirred at room temperature for two hours. The reaction mixture is then poured into water and extracted with an organic solvent, preferably ethyl acetate. The organic extract is washed with water and then with ice-cold 10% aqueous sodium hydroxide. The aqueous alkaline fraction is then acidified, preferably with dilute aqueous hydrochloric acid and extracted with an organic solvent, preferably, chloroform. The chloroform extract is dried, filtered and concentrated under vacuum to give G. The crude residue may further be purified by column chromatography.
The 2-xcex1-acyl hydrocarbylbenzophenone, G (5 g) is dissolved in glacial acetic acid (15 mL). To this mixture is added 60% perchloric acid (7.5 mL). The resulting mixture is warmed to 100xc2x0 C. (steam bath) for three minutes. The mixture is allowed to cool to room temperature. Crystallization of the crude product may begin spontaneously at this point or may be induced by addition to the reaction mixture of ether or ethyl acetate. The product 2-benzopyrylium salt H is removed by filtration and purified by recrystallization, preferably from ethanol or glacial acetic acid/ethyl acetate.
A synthetic sequence, similar to that outlined above, for preparation of 2,3-benzodiazepines is disclosed in U.S. Pat. No. 3,736,31 5, the entire disclosure of which is incorporated herein by reference. Synthetic strategies for preparation of 2,3-benzodiazepines are also disclosed in Horvath et al., Progress in Neurobiology 60(2000) p309-342 and references cited therein; the entire disclosures of which are incorporated herein by reference.
Alternative methods for preparation of intermediate H start with an aryl acetonide or indanone starting material. See Kunnetsov, E. V., and Dorofeenko, G. N., Zh. Org. Khim., 6, 578-581; and M. Vajda, Acia Chem. Acad. Sci. Hung., 40, p.295-307, 1964, respectively. Another variation for preparing 2,3-benzodiazepines is illustrated in Scheme 2 and 3 (Examples 1 and 2). The synthesis proceeds from intermediate G without isolation of the intermediate benzopyrilium salt H.
Resolution of 5-substituted-2,3-benzodiazepines of formula I
The synthetic procedures shown (or referenced) above result in racemic 2,3-benzodiazepines of formula I. The racemate must be resolved in order to isolate the individual (R)- and (S)-enantiomers. Enantiomeric resolution may be achieved by converting racemic compositions of formula I to a pair of diastereomers by either covalently bonding to an optically active moiety, or by salt formation with an optically active base or acid. Either of these two methods provides a molecule with a second chiral center, thus generating a pair of diastereomers. This diastereomeric pair is then separated by conventional methods such as for example, crystallization or chromatography.
Racemic compounds of formula I may be converted to the (S)-dibenzoyltartaric acid salt, which is a diastereomeric mixture of SS and RS configurations. The pair of diastereomers (R,S) and (S,S) possess different properties, e.g., differential solubilities, that allow for the use of conventional separation methods. Fractional crystallization of diastereomeric salts from a suitable solvent is one such separation method. This resolution has been successfully applied to the resolution of racemic tofisopam. See Hungarian Patent 178516 and also Toth et al., J. Heterocyclic Chem., 20:09-713 (1983), the entire disclosures of which are incorporated herein by reference.
Alternatively, racemic compounds of formula I may be derivatized via, for example, acylation of the aromatic hydroxy moiety with a chiral acylating reagent such as, for example, (S)-mandelic acid. The resulting ester, has a second chiral center, and thus exists as a diastereomeric pair separable using conventional methods such as crystallization or chromatography. Following the separation, the chiral moiety with which the compound was derivatized, may be removed.
Racemic compounds of formula I may be separated without diastereomer formation by differential absorption on a chiral stationary phase of a chromatography column, particularly a preparative HPLC column. Chiral HPLC columns are commercially available with a variety of packing materials to suit a broad range of separation applications. Exemplary stationary phases suitable for resolving the racemic 2,3-benzodiazepines include:
(i) macrocyclic glycopeptides, such as silica-bonded vancomycin which contains 18 chiral centers surrounding three pockets or cavities;
(ii) chiral xcex11-acid glycoprotein;
(iii) human serum albumin; and
(iv) cellobiohydrolase (CBH).
Chiral xcex11-acid glycoprotein is a highly stable protein immobilized onto spherical silica particles that tolerates high concentrations of organic solvents, high and low pH, and high temperatures. Human serum albumin, though especially suited for the resolution of weak and strong acids, zwitterionic and nonprotolytic compounds, has been used to resolve basic compounds. CBH is a very stable enzyme that has been immobilized onto spherical silica particles and is preferentially used for the separation of enantiomers of basic drugs from many compound classes.
The resolution of tofisopam by chiral chromatography using macrocyclic glycopeptide as a stationary phase on a Chirobiotic V(trademark) column (ASTEAC, Whippany, N.J.) is disclosed in U.S. Pat. No. 6,080,736. Fitos el al. (J. Chromatogr., 709 265 (1995)), the entire disclosures of which arc incorporated herein by reference, discloses another method for resolving racemic tofisopam by chiral chromatography using a chiral xcex11-acid glycoprotein as a stationary phase on a CHIRAL-AGP(trademark) column (ChromTech, Cheshire, UK). This method separates the (R)- and (S)-enantiomers and also resolves the two conformers (discussed below) of each enantiomer. These methods, may be used to separate racemic 2,3-benzodiazepines of formula I into individual (R)- and (S)-enantiomers. The Chirobiotic V(trademark) column is available in a semi-preparative size as employed for the above separation 500 mmxc3x9710 mm). In addition, the stationary phase of the Chirobiotic V(trademark) column is commercially available in bulk for packing of preparative chromatography columns with larger sample capacity.
In addition to existing as (R)- and (S)-enantiomers, 2,3-benzodiazepines of formula I, also exist in two stable conformations that may be assumed by the benzodiazepine ring as generally depicted below. 
The present invention includes compositions and methods as described herein that use any and all observable conformations of compounds of formula I.
The compound used in the compositions and methods of the present invention may take the form of a pharmaceutically-acceptable salt. The term xe2x80x9csaltsxe2x80x9d, embraces salts commonly used to form alkali metal salts and to form addition salts of free acids or free bases. The term xe2x80x9cpharmaceutically-acceptable saltxe2x80x9d refers to salts that possess toxicity profiles within a range so as to have utility in pharmaceutical applications. Pharmaceutically unacceptable salts may nonetheless possess properties such as high crystallinity, which have utility in the practice of the present invention, such as for example utility in a synthetic process or in the process of resolving enantiomers from a racemic mixture. Suitable pharmaceutically-acceptable acid addition salts may be prepared from an inorganic acid or from an organic acid. Examples of such inorganic acids are hydrochloric, hydrobromic, hydroiodic, nitric, carbonic, sulfuric and phosphoric acid. Appropriate organic acids may be selected from aliphatic, cycloaliphatic, aromatic, araliphatic, heterocyclic, carboxylic and sulfonic classes of organic acids, example of which are formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucuronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, mesylic, salicyclic, salicyclic, 4-hydroxybenzoic, phenylacetic, mandelic, embonic (pamoic), methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, 2-hydroxyethanesulfonic, toluenesulfonic, sulfanilic, cyclohexylaminosulfonic, stearic, algenic, beta-hydroxybutyric, salicyclic, galactaric and galacturonic acid.
Suitable pharmaceutically acceptable base addition salts of compounds of formula I, include for example, metallic salts made from calcium, magnesium, potassium, sodium and zinc or organic salts made from N,Nxe2x80x2-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine. All of these salts may be prepared by conventional means from the corresponding compound of formula I by reacting, for example, the appropriate acid or base with the compound of formula I.
The compounds useful in the methods of the invention may be administered to individuals (mammals, including animals and humans) afflicted with IBS or NUD.
For treating or preventing irritable bowel syndrome or nonulcer dyspepsia, the specific dose of a compound of formula I, or a substantially isolated enantiomer thereof to obtain therapeutic benefit will, of course, be determined by the particular circumstances of the individual patient including, the size, weight, age and sex of the patient. Also determinative will be the nature and stage of the disease and the route of administration. For example, a daily dosage of from about 100 to 1500 mg/kg/day may be utilized. Preferably, a daily dosage of from about 100 to 1000 mg/kg/day may be utilized. More preferably, a daily dosage of from about 100 to 500 mg/kg/day may be utilized. Higher or lower doses are also contemplated.
For prophylactic administration, the compound should be administered far enough in advance of a recurrence of symptoms such that the compound is able to reach the site of action in sufficient concentration to exert a therapeutic effect. The pharmacokinetics of specific compounds may be determined by means known in the art and tissue levels of a compound in a particular individual may be determined by conventional analyses.
The compound may be administered in the form of a pharmaceutical composition comprising at least one compound of formula I in combination with a pharmaceutically acceptable carrier. The active ingredient in such formulations may comprise from 0.1 to 99.99 weight percent. By xe2x80x9cpharmaceutically acceptable carrierxe2x80x9d is meant any carrier, diluent or excipient that is compatible with the other ingredients of the formulation and not deleterious to the recipient.
The compound may be administered for therapeutic effect by any route, for example enteral (e.g., oral, rectal, intranasal, etc.) and parenteral administration. Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intraperitoneal, intravaginal, intravesical (e.g., into the bladder), intradermal, topical or subcutaneous administration. Also contemplated within the scope of the invention is the instillation of drug in the body of the patient in a controlled formulation, with systemic or local release of the drug to occur at a later time. For administration in the therapy of chronic disorders, the compound may optionally be localized in a depot for controlled or sustained release to the circulation, or controlled or sustained release to a local site such as for example the gastrointestinal tract or a portion thereof.
The pharmaceutically acceptable carrier is selected on the basis of the selected route of administration and standard pharmaceutical practice. The active agent may be formulated into dosage forms according to standard practices in the field of pharmaceutical preparations. See Alphonso Gennaro, ed., Remington""s Pharmaceutical Sciences, 18th Ed., (1990) Mack Publishing Co., Easton, Pa. Suitable dosage forms may comprise, for example, tablets, capsules, solutions, parenteral solutions, troches, suppositories, or suspensions.
For parenteral administration, the active agent may be mixed with a suitable carrier or diluent such as water, an oil (particularly a vegetable oil), ethanol, saline solution, aqueous dextrose (glucose) and related sugar solutions, glycerol, or a glycol such as propylene glycol or polyethylene glycol. Solutions for parenteral administration preferably contain a water-soluble salt of the active agent. Stabilizing agents, antioxidizing agents and preservatives may also be added. Suitable antioxidizing agents include sulfite, ascorbic acid, citric acid and its salts, and sodium EDTA. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben, and chlorbutanol. The composition for parenteral administration may take the form of an aqueous or nonaqueous solution, dispersion, suspension or emulsion.
For oral administration, the active agent may be combined with one or more solid inactive ingredients for the preparation of tablets, capsules, pills, powders, granules or other suitable oral dosage forms. For example, the active agent may be combined with at least one excipient such as fillers, binders, humectants, disintegrating agents, solution retarders, absorption accelerators, wetting agents absorbents or lubricating agents. According to one tablet embodiment, the active agent may be combined with carboxymethylcellulose calcium, magnesium stearate, mannitol and starch, and then formed into tablets by conventional tableting methods.
The compositions useful in the methods of the present invention may also be formulated so as to provide slow or controlled-release of the active ingredient therein. In general, a controlled-release preparation is a composition capable of releasing the active ingredient at the required rate to maintain constant pharmacological activity for a desirable period of time. Such dosage forms may provide a supply of a drug to the body during a predetermined period of time and thus maintain drug levels in the therapeutic range for longer periods of time than other non-controlled formulations.
For example, U.S. Pat. No. 5,674,533 discloses controlled-release compositions in liquid dosage forms for the administration of moguisteine, a potent peripheral antitussive. U.S. Pat. No. 5,059,595 describes the controlled-release of active agents by the use of a gastro-resistant tablet for the therapy of organic mental disturbances. U.S. Pat. No. 5, 591,767 discloses a liquid reservoir transdermal patch for the controlled administration of ketorolac, a non-steroidal anti-inflammatory agent with potent analgesic properties. U.S. Pat. No. 5,120,548 discloses a controlled-release drug delivery device comprised of swellable polymers. U.S. Pat. No. 5,073,543 discloses controlled-release formulations containing a trophic factor entrapped by a ganglioside-liposome vehicle. U.S. Pat. No. 5,639,476 discloses a stable solid controlled-release formulation having a coating derived from an aqueous dispersion of a hydrophobic acrylic polymer. The patents cited above are incorporated herein by reference.
Biodegradable microparticles may be used in the controlled-release formulations of this invention. For example, U.S. Pat. No. 5,354,566 discloses a controlled-release powder that contains the active ingredient. U.S. Pat. No. 5,733,566 describes the use of polymeric microparticles that release antiparasitic compositions. These patents are incorporated herein by reference.
The controlled-release of the active ingredient may be stimulated by various inducers, for example pH, temperature, enzymes, water, or other physiological conditions or compounds. Various mechanisms of drug release exist. For example, in one embodiment, the controlled-release component can swell and form porous openings large enough to release the active ingredient after administration to a patient. The term xe2x80x9ccontrolled-release componentxe2x80x9d in the context of the present invention is defined herein as a compound or compounds, such as polymers, polymer matrices, gels, permeable membranes, liposomes and/or microspheres, that facilitate the controlled-release of the compound of formula I in the pharmaceutical composition. In another embodiment, the controlled-release component may be biodegradable, induced by exposure to the aqueous environment, pH, temperature, or enzymes in the body. In another embodiment, sol-gels may be used, wherein the active ingredient is incorporated into a sol-gel matrix that is a solid at room temperature. This matrix is implanted into a patient, preferably a mammal, having a body temperature high enough to induce gel formation of the sol-gel matrix, thereby releasing the active ingredient into the patient.